EGFR signaling plays important roles in the development of eccrine sweat glands. We previously demonstrate that Matrigel induces eccrine sweat gland cells to reconstruct the three-dimensional (3D) structures of eccrine sweat glands, but the mechanisms are still unknown. In the study, eccrine sweat gland cells were cultured within a 3D Matrigel, and EGFR inhibitor AG1478, or MEK1/2 inhibitor U0126, were added to the medium respectively. The morphology of the 3D-reconstructed eccrine sweat gland-like structures was observed, the localization of phospho-EGFR was detected, and protein levels of EGFR, phospho-EGFR, phospho-JAK, phospho-AKT and phospho-ERK were examined. The results showed that cells treatment with AG1478 from Day 0 of 3D cultures blocked formation of spheroid-like structures. AG1478 administration caused reduced phospho-EGFR, concomitant with downregulation of phospho-ERK1/2, but not phospho-JAK or phospho-AKT. Phospho-EGFR and phospho-ERK were reduced, and only a small number of 3D-structures were formed following treatment with U0126. We conclude that EGFR plays important roles in Matrigel-induced 3D structures of eccrine sweat gland-like structures, and ERK1/2 signaling is responsible, at least in part, for the effect of EGFR.

中文翻译：

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EGFR抑制剂AG1478在Matrigel诱导的内分泌汗腺样结构的3D重建中主要通过ERK信号传导途径阻断3D结构的形成。

EGFR信号在内分泌汗腺的发育中起重要作用。我们以前证明Matrigel诱导内分泌汗腺细胞来重建内分泌汗腺的三维（3D）结构，但机制仍是未知的。在这项研究中，内分泌汗腺细胞在3D Matrigel中培养，并将EGFR抑制剂AG1478或MEK1 / 2抑制剂U0126添加到培养基中。观察3D重建的内分泌汗腺样结构的形态，检测磷酸化EGFR的定位，并检测EGFR，磷酸化EGFR，磷酸化JAK，磷酸化AKT和磷酸化ERK的蛋白质水平。结果表明，从3D培养的第0天开始，用AG1478处理细胞可阻止类球体结构的形成。AG1478给药导致磷酸化EGFR降低，伴随磷酸ERK1 / 2，而不磷酸JAK或磷酸AKT的下调。用U0126处理后，磷酸化EGFR和磷酸化ERK减少，仅形成少量3D结构。我们得出结论，EGFR在基质胶诱导的内分泌汗腺样结构的3D结构中起重要作用，而ERK1 / 2信号至少部分负责EGFR的作用。